QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

HOME  |   SEARCH  |   ARCHIVE  |   SUBSCRIBE  |   CONTACT  |   HELP

The Journal of Neuroscience, September 3, 2003, 23(22):8167-8175

This Article Full Text Full Text (PDF) Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (22) Citing Articles via Google Scholar Google Scholar Articles by Zheng, J. Articles by Zagotta, W. N. Search for Related Content PubMed PubMed Citation Articles by Zheng, J. Articles by Zagotta, W. N.

 Previous Article  |  Next Article 

Disruption of an Intersubunit Interaction Underlies Ca²⁺-Calmodulin Modulation of Cyclic Nucleotide-Gated Channels

Jie Zheng,¹ Michael D. Varnum,² and William N. Zagotta¹

¹Howard Hughes Medical Institute and Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, Washington 98195, and ²Washington State University, Department of Veterinary and Comparative Anatomy, Pharmacology, and Physiology, Pullman, Washington 99164

Cyclic nucleotide-gated channels are key molecular elements for olfactory transduction. Olfactory adaptation caused by repeated exposure to an odorant has been proposed to be mediated by the binding of Ca²⁺-calmodulin to the NH₂-terminal domain of the channel, breaking its interaction with the COOH-terminal domain and downregulating the channel. We used a fluorescence resonance energy transfer (FRET) approach to study the structural aspects of this domain-domain interaction under physiological conditions in real time. Fluorescent proteins enhanced cyan fluorescent protein and enhanced yellow fluorescent protein were genetically attached at sites adjacent to the NH₂- and COOH-terminal interacting domains, respectively, allowing direct observation of molecular rearrangements in intact channels. FRET signals caused by the specific interdomain interaction were observed in both intact cells and excised patches. Comparison of the effective FRET efficiencies demonstrated that the interaction occurs specifically between subunits but not within the same subunit. Binding of Ca²⁺-calmodulin caused a reversible decrease in FRET with the same time course as channel downregulation. These results suggest that a separation or reorientation of the interacting domains between subunits by Ca²⁺-calmodulin leads to channel downregulation. The quaternary arrangement presents a structural framework for understanding the molecular mechanism of olfactory adaptation.

Key words: ion channel; Ca²⁺-calmodulin; olfactory adaptation; signal transduction; GFP mutants; FRET; fluorescence

---

Received June 11, 2003; revised July 14, 2003; accepted July 15, 2003.

This article has been cited by other articles:

				C. Waldeck, K. Vocke, N. Ungerer, S. Frings, and F. Mohrlen Activation and desensitization of the olfactory cAMP-gated transduction channel: identification of functional modules J. Gen. Physiol., November 12, 2009; 134(5): 397 - 408. [Abstract] [Full Text] [PDF]

				Z. Qu, W. Cheng, Y. Cui, Y. Cui, and J. Zheng Human Disease-causing Mutations Disrupt an N-C-terminal Interaction and Channel Function of Bestrophin 1 J. Biol. Chem., June 12, 2009; 284(24): 16473 - 16481. [Abstract] [Full Text] [PDF]

				D. P. Mohapatra, D. F. Siino, and J. S. Trimmer Interdomain Cytoplasmic Interactions Govern the Intracellular Trafficking, Gating, and Modulation of the Kv2.1 Channel J. Neurosci., May 7, 2008; 28(19): 4982 - 4994. [Abstract] [Full Text] [PDF]

				M. Pedersen, M. Carmosino, and B. Forbush Intramolecular and Intermolecular Fluorescence Resonance Energy Transfer in Fluorescent Protein-tagged Na-K-Cl Cotransporter (NKCC1): SENSITIVITY TO REGULATORY CONFORMATIONAL CHANGE AND CELL VOLUME J. Biol. Chem., February 1, 2008; 283(5): 2663 - 2674. [Abstract] [Full Text] [PDF]

				D. Willoughby and D. M. F. Cooper Organization and Ca2+ Regulation of Adenylyl Cyclases in cAMP Microdomains Physiol Rev, July 1, 2007; 87(3): 965 - 1010. [Abstract] [Full Text] [PDF]

				J. D. Brady, E. D. Rich, J. R. Martens, J. W. Karpen, M. D. Varnum, and R. L. Brown Interplay between PIP3 and calmodulin regulation of olfactory cyclic nucleotide-gated channels PNAS, October 17, 2006; 103(42): 15635 - 15640. [Abstract] [Full Text] [PDF]

				D. J. D. Espiritu, A. A. Bernardo, and J. A. L. Arruda Role of NH2 and COOH termini in targeting, stability, and activity of sodium bicarbonate cotransporter 1 Am J Physiol Renal Physiol, September 1, 2006; 291(3): F588 - F596. [Abstract] [Full Text] [PDF]

				J. Zheng Patch Fluorometry: Shedding New Light on Ion Channels Physiology, February 1, 2006; 21(1): 6 - 12. [Abstract] [Full Text] [PDF]

				S. Qiu, Y.-l. Hua, F. Yang, Y.-z. Chen, and J.-h. Luo Subunit Assembly of N-Methyl-D-aspartate Receptors Analyzed by Fluorescence Resonance Energy Transfer J. Biol. Chem., July 1, 2005; 280(26): 24923 - 24930. [Abstract] [Full Text] [PDF]

				J. A. Fisher, G. Girdler, and B. S. Khakh Time-Resolved Measurement of State-Specific P2X2 Ion Channel Cytosolic Gating Motions J. Neurosci., November 17, 2004; 24(46): 10475 - 10487. [Abstract] [Full Text] [PDF]

				M. C. Trudeau and W. N. Zagotta Dynamics of Ca2+-Calmodulin-dependent Inhibition of Rod Cyclic Nucleotide-gated Channels Measured by Patch-clamp Fluorometry J. Gen. Physiol., August 30, 2004; 124(3): 211 - 223. [Abstract] [Full Text] [PDF]

Home  |   Search  |   Archive  |   Subscribe  |   Contact  |   Help